File Download

There are no files associated with this item.

  Links for fulltext
     (May Require Subscription)
Supplementary

Article: Potent gene-specific inhibitory properties of mixed-backbone antisense oligonucleotides comprised of 2′-deoxy-2′-fluoro-D-arabinose and 2′-deoxyribose nucleotides

TitlePotent gene-specific inhibitory properties of mixed-backbone antisense oligonucleotides comprised of 2′-deoxy-2′-fluoro-D-arabinose and 2′-deoxyribose nucleotides
Authors
Issue Date2002
PublisherAmerican Chemical Society. The Journal's web site is located at http://pubs.acs.org/biochemistry
Citation
Biochemistry, 2002, v. 41 n. 10, p. 3457-3467 How to Cite?
AbstractPhosphorothioate deoxyribonucleotides (PS-DNA) are among the most widely used antisense inhibitors. PS-DNA exhibits desirable properties such as enhanced nuclease resistance, improved bioavailability, and the ability to induce RNase H mediated degradation of target RNA. Unfortunately, PS-DNA possesses a relatively low binding affinity for target RNA that impacts on its potency in antisense applications. We recently showed that phosphodiester-linked oligonucleotides comprised of 2′-deoxy-2′-fluoro-D-arabinonucleic acid (FANA) exhibit both high binding affinity for target RNA and the ability to elicit RNase H degradation of target RNA [Damha et al. (1998) J. Am. Chem. Soc. 120, 12976]. In the present study, we evaluated the antisense activity of phosphorothioate-linked FANA oligonucleotides (PS-FANA). Oligonucleotides comprised entirely of PS-FANA were somewhat less efficient in directing RNase H cleavage of target RNA as compared to their phosphorothioate-linked DNA counterparts, and showed only weak antisense inhibition of cellular target expression. However, mixed-backbone oligomers comprised of PS-FANA flanking a central core of PS-DNA were found to possess potent antisense activity, inhibiting specific cellular gene expression with EC 50 values of less than 5 nM. This inhibition was a true antisense effect, as indicated by the dose-dependent decrease in both target protein and target mRNA. Furthermore, the appearance of mRNA fragments was consistent with RNase H mediated cleavage of the mRNA target. We also compared a series of PS-[FANA-DNA-FANA] mixed-backbone oligomers of varying PS-DNA core sizes with the corresponding 2′-O-methyl oligonucleotide chimeras, i.e., PS-[2′meRNA-DNA-2′meRNA]. Both types of oligomers showed very similar binding affinities toward target RNA. However, the antisense potency of the 2′-O-methyl chimeric compounds was dramatically attenuated with decreasing DNA core size, whereas that of the 2′-fluoroarabino compounds was essentially unaffected. Indeed, a PS-FANA oligomer containing a single deoxyribonucleotide residue core retained significant antisense activity. These findings correlated exactly with the ability of the various chimeric antisense molecules to elicit RNase H degradation of the target RNA in vitro, and suggest that this mode of inhibition is likely the most important determinant for potent antisense activity.
Persistent Identifierhttp://hdl.handle.net/10722/167752
ISSN
2015 Impact Factor: 2.876
2015 SCImago Journal Rankings: 1.769
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorLok, CNen_US
dc.contributor.authorViazovkina, Een_US
dc.contributor.authorMin, KLen_US
dc.contributor.authorNagy, Een_US
dc.contributor.authorWilds, CJen_US
dc.contributor.authorDamha, MJen_US
dc.contributor.authorParniak, MAen_US
dc.date.accessioned2012-10-08T03:11:02Z-
dc.date.available2012-10-08T03:11:02Z-
dc.date.issued2002en_US
dc.identifier.citationBiochemistry, 2002, v. 41 n. 10, p. 3457-3467en_US
dc.identifier.issn0006-2960en_US
dc.identifier.urihttp://hdl.handle.net/10722/167752-
dc.description.abstractPhosphorothioate deoxyribonucleotides (PS-DNA) are among the most widely used antisense inhibitors. PS-DNA exhibits desirable properties such as enhanced nuclease resistance, improved bioavailability, and the ability to induce RNase H mediated degradation of target RNA. Unfortunately, PS-DNA possesses a relatively low binding affinity for target RNA that impacts on its potency in antisense applications. We recently showed that phosphodiester-linked oligonucleotides comprised of 2′-deoxy-2′-fluoro-D-arabinonucleic acid (FANA) exhibit both high binding affinity for target RNA and the ability to elicit RNase H degradation of target RNA [Damha et al. (1998) J. Am. Chem. Soc. 120, 12976]. In the present study, we evaluated the antisense activity of phosphorothioate-linked FANA oligonucleotides (PS-FANA). Oligonucleotides comprised entirely of PS-FANA were somewhat less efficient in directing RNase H cleavage of target RNA as compared to their phosphorothioate-linked DNA counterparts, and showed only weak antisense inhibition of cellular target expression. However, mixed-backbone oligomers comprised of PS-FANA flanking a central core of PS-DNA were found to possess potent antisense activity, inhibiting specific cellular gene expression with EC 50 values of less than 5 nM. This inhibition was a true antisense effect, as indicated by the dose-dependent decrease in both target protein and target mRNA. Furthermore, the appearance of mRNA fragments was consistent with RNase H mediated cleavage of the mRNA target. We also compared a series of PS-[FANA-DNA-FANA] mixed-backbone oligomers of varying PS-DNA core sizes with the corresponding 2′-O-methyl oligonucleotide chimeras, i.e., PS-[2′meRNA-DNA-2′meRNA]. Both types of oligomers showed very similar binding affinities toward target RNA. However, the antisense potency of the 2′-O-methyl chimeric compounds was dramatically attenuated with decreasing DNA core size, whereas that of the 2′-fluoroarabino compounds was essentially unaffected. Indeed, a PS-FANA oligomer containing a single deoxyribonucleotide residue core retained significant antisense activity. These findings correlated exactly with the ability of the various chimeric antisense molecules to elicit RNase H degradation of the target RNA in vitro, and suggest that this mode of inhibition is likely the most important determinant for potent antisense activity.en_US
dc.languageengen_US
dc.publisherAmerican Chemical Society. The Journal's web site is located at http://pubs.acs.org/biochemistryen_US
dc.relation.ispartofBiochemistryen_US
dc.subject.meshArabinose - Analogs & Derivatives - Chemistryen_US
dc.subject.meshBase Sequenceen_US
dc.subject.meshCircular Dichroismen_US
dc.subject.meshDeoxyribose - Chemistryen_US
dc.subject.meshEscherichia Coli - Enzymologyen_US
dc.subject.meshLuciferases - Metabolismen_US
dc.subject.meshNucleic Acid Conformationen_US
dc.subject.meshOligonucleotides, Antisense - Chemistry - Pharmacologyen_US
dc.subject.meshRibonuclease H - Isolation & Purification - Metabolismen_US
dc.titlePotent gene-specific inhibitory properties of mixed-backbone antisense oligonucleotides comprised of 2′-deoxy-2′-fluoro-D-arabinose and 2′-deoxyribose nucleotidesen_US
dc.typeArticleen_US
dc.identifier.emailLok, CN:cnlok@hku.hken_US
dc.identifier.authorityLok, CN=rp00752en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1021/bi0115075en_US
dc.identifier.pmid11876654-
dc.identifier.scopuseid_2-s2.0-0037066105en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0037066105&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume41en_US
dc.identifier.issue10en_US
dc.identifier.spage3457en_US
dc.identifier.epage3467en_US
dc.identifier.isiWOS:000174387500020-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridLok, CN=7006410829en_US
dc.identifier.scopusauthoridViazovkina, E=6507963057en_US
dc.identifier.scopusauthoridMin, KL=7201466717en_US
dc.identifier.scopusauthoridNagy, E=35345096200en_US
dc.identifier.scopusauthoridWilds, CJ=6701808160en_US
dc.identifier.scopusauthoridDamha, MJ=7007107329en_US
dc.identifier.scopusauthoridParniak, MA=7006356357en_US

Export via OAI-PMH Interface in XML Formats


OR


Export to Other Non-XML Formats